transformed clone 中文意思是什麼

transformed clone 解釋
轉化無性系,轉化克隆
  • transformed : 使. . . 變形
  • clone : n. 1. 【生物學】純種細胞，無性系。2. 復製品；（幾乎）一模一樣的人。3. （不動腦筋）機械行事的人；機器人。vt. 把…培養為純種細胞；無性繁殖；〈比喻〉復制。
  1. Sub - - clone of s, . / hbsag fusion gene : pbuescripts, . / hbsag and ppiczaa were digested separately by xhoi and xbai enzyme, and were linked under t4 dna ligase, ppiczaa s, / hbsag was constructed and transformed to e. coli

    Hbsag質粒與ppiczaa載體分別經xhol和xbaln切，再在t4dna連接酶作用下進行連接，獲得工程菌表達型ppiczaas ； hbsag質粒，轉化大腸桿菌t0p10細胞，經xhol和xbal與sacll和xbal酶切電泳，證實s ； 。
  2. Then the linked products were transformed into the high competent cell of e. coli dh5a. based on - complementation of the detective - galactosidase, positive recombinant clone were screened from x - gal plate

    從引物和基因序列的比對分析結果看， zhyf006序列與上下游引物的配對比例分別為s4
  3. In this study, we designed a pair of primers based on the sequence of the upstream and downstream of chicken il - 2 gene. about 600 bp chicken il - 2 cdna fragment was cloned from cona - stimulated chicken splenocytes by reverse transcription - polymerase chain reaction ( rt - pcr ) and was subcloned into puc18 vector. recombinant clone was demonstrated by restriction enzyme digestion and dna sequencing. next, we construct recombinant plasmid pproex ? t - il - 2. the cdna of chicken il - 2 gene was subcloned into bamh i / hind iii sites of vector. the recombinant plasmid pproex ? t - il - 2 was transformed into e. coli dh5a and the bacteria was induced with iptg. it was demonstrated by sds - page and western blot that a 18kda protein which was equal to chicken il - 2 protein in molecular weight was expressed in e. coli dh5a. the expression level was up to 30 % of the total bacterial proteins. the purified protein was used to prepare the antibody against chicken il - 2 protein

    經酶切鑒定及dna序列測定，該基因為雞il - 2基因，其序列與sundick等報道的完全一致。在此基礎上，我們把雞il - 2基因亞克隆到大腸桿菌原核表達載體pproex ~ （ tm ） ht中，構建重組表達質粒並進行確證性序列測定，重組質粒測序結果表明將編碼雞il - 2成熟蛋白的基因正確地插入到原核表達載體pproex ~ （ tm ） ht的目的位點。重組質粒轉化受體菌dh5後用iptg於37進行誘導培養， sds - page和westernblot分析顯示，表達的雞il - 2融合蛋白分子量約為18kda ，表達的融合蛋白經薄層掃描發現目的蛋白表達量約占菌體蛋白的30 。
  4. Selected one of the 14 strains - s93, s93 dna was digested partially with sau3a i and 2 ~ 3kb fragments were collected and inserted into puc 18, then transformed into dh5 a. filtering the clone with hybridization in situ, a 1 kb frament clone has been cloned

    使用sau3ai對基因組dna進行不完全酶切，回收2 3kb片段，與puc18質粒連接轉化大腸桿菌，利用地高辛標記探針，使用菌落原位雜交篩選轉化子；篩選到包含有約1kb外源片段的轉化子。
  5. After electrophorised on 1 % agarose gel, the pcr production was purified with agarose gel dna extraction kit. the segment was ligated with vector pmd18 - t and then was tranformed into the competent cell of dh5 a. a construction mstnd - pmd18t was generated by inserting the sequence of 254bp into pmd18 - t vector and selecting the sense clones. positive clone was identified by three ways : endonuclease digestion, pcr and sequencing. the result showed that the cloned sequence coincides with the designed sequence. this construction was digested with nco i and xho i and ligated the pet28a ( + ) vector digested with the same enzymes using dna ligation kit. the production of ligation reaction was transformed into the competent cell of bl21 ( de3 ). after 12 - 16 hours of culture, several colnes appeared on the plate. some positive clones were selected to extract their plasmid. these plasmids were digested by nco i and xho i and indentified by pcr. a contraction, mstnd - pet28a was generated. the result showed that the cloned sequence coincides with the designed sequence

    F _ 1長38bp ， r _ 1長36bp ，其它片段均40bp長， f _ 1和r _ 1片段兩端分別加上限制性內切酶nco和xho的識別位點序列。用成對單鏈片段進行延伸反應，然後用其他單鏈片段作為引物，進行pcr擴增，用dna快速純化回收試劑盒回收所得254bppcr產物，與pmd18 - t載體連接、轉化dh _ 5 。受體菌感受態細胞，利用藍白斑遺傳學篩選法篩選陽性克隆，提取其質粒，採用nco和xho雙酶切鑒定，獲得了254bp的片段；用pmd18 - t載體上的特異引物rv - m和m13 - 47進行pcr鑒定，獲得300bp的片段。
  6. By cheeking the transformed bacterial colonies, we had filtrated the masculine clone successfully. the sequencing result showed that the inserting fragment contained intact coding sequences of dh. pban ( pheromone biosynthesis activating neuropeptide ) and three sgnps ( suboesophageal ganglion neuropeptide ) and the leading frame of it was correct

    測序結果表明，重組載體的插入片段中含有dh 、性信息素生物合成激活肽（ pheromonebiosynthesisactivatingneuropeptide ， pban ）及三個食道下神經節肽（ suboesophagealganglionneuropeptide ， sgnp ）的完整的編碼序列，且其閱讀框架（ readingframe ）完全正確。
  7. Main works and their important results are showed as following. firstly, the cdna encoding bullfrog gh is amplified by use of the total rna, extracted from adult bullfrog ' s pituitary, as the template, pi ( 5 ' - cggatcc atg gct tca ggg tta ggc ) and p2 ( 5 ' - cgaattc tta aaa ggt gca gtt gct ) as the primer pair and one particular cdna product, 660bp, was obtained after the rt - pcr - the product was purified by using dna agarose gel extraction method. after the purified cdna and pmd18 - t vector were ligated at 16 over night, the ligation products were transformed into e. coli strain dh5a and then the transformants were screened by clone pcr method

    首先，以成體牛蛙的腦垂體總rna為模板，進行rt ? pcr擴增得到約660bp的特異性pcr產物帶，切下瓊脂糖凝膠中的特異產物帶進行cdna膠回收，將cdna與pmd18 ? t載體進行t ? a克隆連接並轉化到dh5 e . coli后，進行菌落pcr 、質粒酶切鑒定，篩選出陽性菌株，測序結果經blast分析，與已報道的牛蛙生長激素基因高度同源，證實此陽性克隆為牛蛙生長激素基因轉化子，命名為pbfgh 。
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